Metabolism effector links in diet-induced and genetically-based obesity: A full-transcriptome study of liver tissue in experimental models in rodents

Background. When developing methods for personalized diet therapy of obesity, an urgent task is to study the molecular genetics features of the obesity pathogenesis using in vivo experimental models in laboratory animals.The aim. To determine metabolism effector links in obesity based on a comparative analysis of full-transcriptome profiles of the liver tissue of mice and rats of various strains.Materials and methods. We carried out a comparative analysis of the changes in liver transcriptome in rats and mice fed with diets of excessive energy value and exerting lipogenic effect. Data of full-transcriptome profiling using DNA microarray technology have been presented previously in 8 publications. Results. In three strains of mice treated with a high-carbohydrate high-fat diet (HCHFD), a significant differential expression (DE) of 1849 genes was revealed, of which 74 genes responded jointly in at least two groups of animals. In Wistar and Zuckerfa rats, 2109 genes responded to the consumption of HCHFD, of which 242 genes responded jointly in two groups of animals. For rodents different in genetic predisposition to the development of diet-induced obesity, the groups of genes that responded with the opposite sign of DE (depending on the genotype) in reaction to the consumption of HCHFD were identified. Bioinformatical analysis allowed establishing the presence of 43 metabolic pathways, which are targeted for the applied experimental diets exposure, in rats and 77 pathways – in mice. Four of these pathways – the pathway of retinoid metabolism, PPAR signaling pathway associated with it the previous one, xenobiotics metabolism and drugs metabolism mediated by cytochrome P450 system – responded in all groups of animals (except for female mice). The importance of the expression of Tat gene encoding tyrosine aminotransferase in the modulation of biogenic amines synthesis in diet-induced obesity was shown, which may represent a new neurometabolic regulatory function of the liver in response to the consumption of high-calorie diets. Conclusion. The analysis of the results of full-transcriptome studies showed that within each studied species (Rattus rattus and Mus domesticus) and animal sex, a number of genetic variants with a greater or lesser predisposition to the development of diet-induced obesity phenotype can be identified; and at the same time, within these variants, there is a largely similar pattern in the response of metabolism effector links to hypercaloric dietary intake. This pattern creates new prospects for translating the results of transcriptomic and metabolomic studies of laboratory animals into clinical practice in order to substantiate new approaches to personalized diet therapy of alimentary dependent diseases in patients with different genetic predisposition to obesity

Interaction of bacterial extracellular microvesicles with eukaryotic cells.

Bacterial extracellular microvesicles (BMV) are formed by nonpathogenic, pathogenic and opportunistic bacteria. BMV are spherical bilayer-membrane organelles containing different cargoes: lipopolysaccharides, pathogen associated molecular patterns (PUMP), DNA, RNA, signal molecules, proteins, antibiotic resistance factors, virulence factors, toxins providing various immune response options and conducive to the survival and pathogen dissemination in the human body. BMVs secretion play an important role in the ability of microorganisms to cause various diseases. BMV are involved in biofilms formation, help bacteria to obtain nutrition in a nutrient-poor conditions, to evade the host's immune response, provide communication and surviving in a stressful environment during infection inside the host. The heterogeneity of the biogenesis mechanisms causes differences in the BMV and their characteristics including virulence rate. BMVs host cells entering is mediated by several mechanisms and helps to activate innate and adaptive immune reactions. This review focuses on interaction study of BMV with various eukaryotic cells types including neutrophils, dendritic cells, macrophages, epithelial, endothelial cells. This interaction depends on bacteria species, type of target cell and number of vesicles and can lead to different responses: non-immunogenic, pro-inflammatory, cytotoxic. Subcellular and molecular mechanisms related to the involvement of extracellular microvesicles in host's immune response modulation are presented. Stimulation of immune response is provided by increased secretion of proinflammatory cytokines and chemokines. In some cases BMV use mechanisms to evade immune surveillance: anti-inflammatory cytokines secretion, alterations of phagocytosis and chemotaxis of macrophages, increasing the proteolytic cleavage of CD14 on the macrophage surface, alterations of antigen-presenting function of dendritic cells, T-cell proliferation suppression, reducing the pro-inflammatory cytokines secretion, evasion of host-immune cells direct interactions, destruction of neutrophilic traps. These features allow bacterial cells to survive in the human body, increase their invasive potential, and reduce the excessive inflammatory reactions leading to death of the pathogen itself and life-threatening damage of tissues and organs of the host. Further studies of these mechanisms will improve existing therapeutic approaches to the infectious diseases treatment

Anthocyanins as a factor in the alimentary restoration of cellular immunity in diet induced obesity in rats

The article presents the results of a study of the effect of anthocyanins on cellular immunity in rats on a model of alimentary obesity. The aim of the study was to study the effect of an anthocyanin- enriched diet on cellular immunity in diet induced obesity in rats. The study was carried out on male Wistar rats with an initial body weight of 108±2 g. The animals were randomized by body weight into 3 groups (8 pcs. in group). For 12 weeks, rats of the 1st (control) group received a complete modified diet of AIN93M; rats of the 2nd group consumed a high-calorie choline-deficient diet (HCChDD), the fat content of which was 45%, fructose – 20% of the energy value of the diet; rats of the 3rd group received HCChDD with the addition of standardized blueberry and blackcurrant extract (30% anthocyanins) at an average daily dose of 11 mg anthocyanins/kg body weight. The expression of differentiation markers of peripheral blood lymphocytes was carried out by flow cytofluorimetry. As a result of the study, it was found that in rats of the 2nd group with alimentary obesity, the relative content in the peripheral blood of T helpers (CD3+CD4+) was increased (p < 0.05) (75.75±1.11% versus 70.07±0 49% – group 1, 72.14±0.91% – group 3) and reduced (p < 0.05) content of T cytotoxic lymphocytes (CD3+CD8+) (22.54±1.14% versus 28.09±0.72% – 1st group, 26.07±0.87% – 3rd group). The CD3/CD4 ratio in rats of the 2nd group exceeded (p < 0.05) this index in rats of the 1st and 3rd groups (3.44±0.25 versus 2.47±0.09 – 1st group, 2.79±0.13 – 3rd group). Enrichment of the HCChDD with the blueberry and blackcurrant extract led to the normalization of these parameters of cellular immunity. The number of B lymphocytes (CD45R+), Т lymphocytes (CD3+) and NK cells (CD161+) in the rat peripheral blood of all experimental groups had no statistically significant differences. The results of the study of cellular immunity in rats with alimentary obesity indicate the presence of metainflammation. The received data indicate the prospect of using biologically active substances

Prevalence of Listeria monocytogenes in meat products during 2017–2019 depending on technological factors and seasons

Microbiological examination of contamination of imported and domestic meat products with pathogenic bacteria Listeria monocytogenes depending on a meat type, technology and season was carried out during 2017–2019. In total, 2777 product samples were analyzed; the presence of this pathogen was revealed in 8.8% of products (244 positive samples). It was found that the prevalence of L. monocytogenes in meat products increased over three years of observation (2017–2019). The highest occurrence of this pathogen was found in poultry meat (on average 18.7%) followed by products from beef (13.2%). Meat products from mixed raw materials (beef and pork) accounted for 5.3% of tested samples, while in pork semi-finished products L. monocytogenes was found only in 3.2% of cases. It was noted that the technology of semi-finished products significantly affected the level of contamination of meat products with L. monocytogenes. Various technological approaches are used in the production process increasing the risk of contamination of the finished product since there is no timely data on Listeria contamination of raw materials used for production of a particular product. It has been established that a significant role in microbiological studies is played by various approaches to sample preparation of analyzed samples of meat cuts, semi-finished products in large and small pieces, as well as minced semi-finished products. Not knowing the real level of surface contamination with L. monocytogenes of carcasses, half-carcasses, semi-finished products in large pieces, manufacturers use such raw materials for the subsequent production of other types of semi-finished meat products, increasing the risk of manufacturing unsafe products with following contamination of equipment, work surfaces and other objects of the production environment. The highest occurrence of L. monocytogenes in meat products during three years of observation was found in the summer period (14.2%). The proportions of positive samples in the winter, spring and autumn months varied on average within 6.7–7.1%

Evaluation of the regulatory effect of carnosine and alpha-lipoic acid on the cytokine profile of the cerebral cortex of Wistar rats under induced obesity

BACKGROUND: The model of obesity under experimental conditions is reproduced by using high-calorie diets in animals. It has been established that metabolic disorders cause meta-inflammation not only in peripheral organs and tissues, but also in brain structures. The search for effective neuroprotective antioxidants to suppress inflammatory processes in the cerebral cortex in obesity is an urgent task due to the widespread prevalence of this disease.AIM: to evaluate the effect of minor biologically active substances — carnosine (CAR) and α-lipoic acid (ALA) on the cytokine profile of the frontal cortex of the left hemisphere of the brain in Wistar male rats with obesity induced by a high-calorie choline-deficient diet.MATERIALS AND METHODS: The studies were carried out on male Wistar rats with an initial body weight of 150±10 g. The animals were randomized by body weight into 5 groups. For 8 weeks, rats of the 1st (control) group received a complete modified diet of AIN93M; rats of the 2nd group consumed a high-calorie choline-deficient diet (HCHDR), the fat content of which was 45%, fructose — 20% of the energy value of the diet; rats of the 3rd group received HCHDR with the addition of CAR at a dose of 75 mg per 1 kg of body weight; rats of the 4th group received HCHDR with the addition of ALA at a dose of 75 mg per 1 kg of body weight; rats of the 5th group received HCHDR with the addition of the CAR + ALA complex in a total dose of 150 mg per 1 kg of body weight. Animals were removed from the experiment by decapitation under ether anesthesia. The levels of triglycerides (Tg) and free fatty acids (FFA) in blood plasma (mmol) were determined on a biochemical analyzer (Konelab 20i, Thermo Clinical Labsystems Oy, Finland). Content of cytokines and chemokines (pg/ml): GM-CSF, IL-10, IL-17A, IL-12p40, IL-12p70, IL-1α, IL-2, IL-4, IL-5, IFN-γ, MCP-1, M-CSF, MIP-1α, MIP-2, MIP-3α, RANTES, and TNF-α in cerebral cortex lysates were determined by multiplex immunoassay using a Luminex 200 analyzer (Luminex Corporation, USA). To assess the relationship between the level of cytokines in blood plasma and changes in their concentrations under the influence of HCCDR in lysates of the cortex of the frontal lobe of the left hemisphere of the brain, the ratio was calculated: the level of cytokines pg/ml in blood plasma [1]/the content of cytokines pg/ml in lysates (pl/ lys) for each sample.RESULTS: On the model of obesity in rats, the presence of an inflammatory process in the cerebral cortex was established, as evidenced by an increase in the content of pro-inflammatory factors: IL-2, M-CSF, MIP-1α and RANTES and a decrease in the content of immunoregulatory cytokines of varying severity: IL-10, IL17A, IL-12p40, IL-12p70, TNF-a, MIP-2 and MIP-3α in group 2 rats. (HCHDR) compared with the control group. Enrichment of HCHDR with biologically active substances: CAR, ALA or their complex, ensured the normalization of lipid metabolism, as evidenced by the decrease in the ratio of circulating Tg to FFA in the blood serum of rats to control values: 1st gr. (control) — 1,04±0.23; 2nd gr. (HCHDR) — 1,64±0.63; 3rd gr. (CAR) — 0,98±0.31; 4th gr. (ALA) — 0,86±0.31; 5th gr. (CAR+ALA) — 1,02±0.38. Enrichment of HCHDR with CAR, ALA or their complex led to a decrease in the content of pro-inflammatory and apoptosis-regulating cytokines and chemokines in the cortex of the frontal lobe of the rat brain: IL-1α, IL-2, IL-17A, M-CSF, MCP-1, MIP3α and RANTES, along with an increase in the level of the anti-inflammatory cytokine IL-10, which indicates the suppression of the inflammatory process induced by the consumption of HCHDR in rats.CONCLUSION: The data obtained indicate the prospect of using CAR and ALA or their complex as neuroprotective antioxidants to reduce the inflammatory process in brain structures in obesity

КЛИНИКО-ИММУНОЛОГИЧЕСКОЕ ОБОСНОВАНИЕ ПРИМЕНЕНИЯ ВИТАМИННО-МИНЕРАЛЬНОГО КОМПЛЕКСА И ИМУНОФАНА У РЕКОНВАЛЕСЦЕНТОВ ПОСЛЕ ГЕМОРРАГИЧЕСКОЙ ЛИХОРАДКИ С ПОЧЕЧНЫМ СИНДРОМОМ

The efficiency of using Imunophan, Milgamma, Vitrum-antioxidant in a multimodality therapy during a recovery period for patients with hemorrhagic fever with renal syndrome has been studied. Statistically-valid the given therapy leads to the decrease of TNF-α, IL-4, IL-10 level, the increase of INF-γ и IL-2 levels, improves cellular immunity and increases an induction index and working balance of phagocytes. Stabilization of vitamin status was exposed with patients suffering from a middle form of hemorrhagic fever with renal syndrome (HFRS) and significant improvement of a vitamin ratio with patients suffering from a bad form of HFRS. Positive influence on phylaxis , vitamin status, an essentialmicroelement level leads to a clinical improvement of disease state.Изучена эффективность Имунофана, Мильгамы, Витрум-антиоксиданта в комплексной терапии реконвалесцентного периода геморрагической лихорадки с почечным синдромом. Выявлено, что данная терапия способствует статистически достоверному уменьшению уровня TNF-α, IL-4, IL-10 и повышению уровней INF-γ и IL-2, оказывает нормализующее действие на показатели клеточного иммунитета, увеличивает ёмкость функционального резерва фагоцитов. Отмечается нормализация витаминного статуса при среднетяжелой форме геморрагической лихорадки с почечным синдромом (ГЛПС) и значительное улучшение показателей обеспеченности витаминами при тяжелой форме ГЛПС. Положительное влияние на иммунитет, витаминный статус, содержание эссенциальных микроэлементов способствует клиническому улучшению течения заболевания

Прогностическое значение генетического полиморфизма промоторной области AQP5 при сепсисе с различными очагами

Aquaporins represent proteins contributed to water transport through cell membrane. They are involved in formation and resolution of edema, cell migration and inflammatory reaction. There are only few studies linking the genetic polymorphism of aquaporin 5 (rs3759129 AQP5) and sepsis. At the same time, the apparent heterogeneity of patients along the foci of infection may limit finding the most significant association of AQP5 genotypes with the course of infectious complications of critical conditions and restrict further development of rs3759129 AQP5 as a potentially strong marker of sepsis outcome.The purpose of the study was to determine whether the preferential localization of the infection affects the prognostic value of the genetic marker AQP5 (1364A/C, rs3759129) in outcome prediction in sepsis (SEPSIS-3, 2016) patients.Materials and methods. Study groups (n=339) included ICU patients with abdominal sepsis (AS, including pancreatitits, peritonitis, cholecystitis, appendicitis; n=94) sepsis patients with other sources of infections  (n=65) and ICU patients without sepsis (n=180). AQP5 polymorphism was studied by analyzing PCR products in a 2% agarose gel using a AQP5 1364A/C specific tetra primer set.Result. Distribution of alleles (A and C) and genotypes (AA, AC and CC) AQP5 1364A/C in patients with  sepsis or sepsis subgroups (sepsis with no septic shock and sepsis shock patients) versus control group (healthy  volunteers) did not differ. Although there was a trend to preferential survival of sepsis patients with genotype C AQP5 despite the source of infection, only patients with AQP5 CC or AC genotype and abdominal sepsis (Sepsis-3), or a subgroup of the same AQP5 genotype experiencing septic shock, demonstrated increased 30day survival versus AA homozygotic patients (P=0.002).Conclusion. The informative value of detecting the AQP5 CC or AC genotype for prognosis of 30-day survival versus AA homozygotic patients is most significant only in abdominal sepsis patients.Аквапорины — мембранные белки, играющие роль в транспорте молекул воды через клеточную мембрану и участвующие в формировании и разрешении отеков, миграции клеток, воспалительных реакциях. Имеются единичные исследования, свидетельствующие о связи генетического полиморфизма аквапорина 5 (rs3759129 AQP5) с течением сепсиса. Вместе с тем, очевидная гетерогенность пациентов по очагам инфекции может затруднить поиск наиболее выраженной ассоциации генотипов AQP5 с течением инфекционных осложнений критических состояний и дальнейшую разработку rs3759129 AQP5 как потенциально сильного маркера исхода сепсиса.Цель исследования: выяснить связь аллельных вариантов сайта однонуклеотидного полиморфизма гена AQP5 (1364A/C, rs3759129) с исходами сепсиса (СЕПСИС-3, 2016) в зависимости от вероятного первичного очага инфекции.Материалы и методы. С помощью тетрапраймерной полимеразной цепной реакции с последующей электрофоретической визуализацией продуктов проведено аллель-специфическое генотипирование ДНК, выделенной из образцов крови 339 пациентов отделений реанимации и интенсивной терапии двух лечебных учреждений.Результаты. Выявлена тенденция к преимущественному выживанию пациентов с сепсисом с генотипами AQP5 «С+» (AС и CC) вне зависимости от источника инфекции (p>0,050). Однако только в группе пациентов с генотипами AQP5 AC или CC и абдоминальным сепсисом (Sepsis-3, 2016) было выявлено значительное увеличение 30-дневной выживаемости по сравнению с гомозиготными пациентами генотипа AQP5 АА (p=0,002).Заключение. Информативная ценность выявления генотипов CC или AC AQP5 для прогноза 30дневной выживаемости по сравнению с гомозиготными пациентами с генотипом AA может быть выше у пациентов с абдоминальным сепсисом

АЛЛЕЛЬНЫЕ ВАРИАНТЫ ГЕНОВ NRF2 И TLR9 ПРИ КРИТИЧЕСКИХ СОСТОЯНИЯХ

Aim of the study. To elucidate the association of allelic variants of single nucleotide polymorphism in NRF2 (rs6726395, 177238501A>G) and TLR9 (rs352162, 52218953T>C) genes, each gene separately and in their combination, with peculiarities of the course of critical conditions during lung infection. Materials and methods. DNA from 86 post#operative patients and oncologic patients was genotyped in an allelespecific fashion using tetra#primer polymerase chain reaction followed by gel electrophoresis analysis of products.Results. It has been found that septic shock patients with NRF2 177238501A>G GG genotype had increased mortality and higher APACHE II score and developed non#responsive edema more frequently. Patients with NRF2 177238501A>G GG/TLR9 52218953T>C CC genotype combination developed septic shock and nosocomialpneumonia more rarely.Conclusion. The homozygous NRF2 177238501A>G (GG) allele combination is unfavorable for the course and outcome of critical conditions only in combination with TLR9 52218953T>C СT or TLR9 52218953T>C TT alleles in septic shock patients. At the same time, the combination of TLR9 52218953T>C СС alleles in the same patients with 'unfavorable' NRF2 177238501A>G GG protects against development of septic shock and nosocomial pneumonia.Цель исследования. Выявить связь аллельных вариантов сайтов однонуклеотидного полиморфизма генов NRF2 (rs6726395, 177238501A>G) и TLR9 (rs352162, 52218953T>C) — изолированно и в комплексе — с особенностями течения критических состояний при инфекции легких. Материалы и методы. Аллель#специфически генотипирована ДНК от 86 больных, перенесших хирургическое вмешательство, и онкологических больных.Результаты. Обнаружено, что у больных генотипа NRF2 177238501A>G GG с септическим шоком повышена летальность и более высокие баллы по шкале APACHE II, чаще развивался отек легких, резистентный к лечению. У больных с комбинацией генотипов NRF2 177238501A>G GG/TLR9 52218953T>C CC реже развивался септический шок и нозокомиальная пневмония.Заключение. Комбинация аллеля NRF2 177238501A>G в гомозиготном состоянии (GG) является неблагоприятной для течения и исхода критических состояний только в сочетании с аллелями TLR9 52218953T>C СT или TLR9 52218953T>C TT у больных с септическим шоком. При этом комбинация аллелей TLR9 52218953T>C СС с «неблагоприятными» NRF2 177238501A>G GG защищает от развития септи#ческого шока и нозокомиальной пневмонии

Biomarkers of Air-Blood Barrier Damage In COVID-19

The search for sensitive and specific markers enabling timely identification of patients with a life-threatening novel coronavirus infection (COVID-19) is important for a successful treatment.The aim of the study was to examine the association of molecular biomarkers of air-blood barrier damage, surfactant proteins SP-A and SP-D and Club cell protein CC16, with the outcome of patients with COVID-19.Materials and methods. A cohort of 109 patients diagnosed with COVID-19 was retrospectively divided into two groups. Group 1 comprised survivor patients discharged from the ICU (w=90). Group 2 included the patients who did not survive (w=19). Association of disease outcome and SP-A, SP-D, and CC16 levels in blood serum, clinical, and laboratory data were examined taking into account the day of illness at the time of biomaterial collection.Results. The non-survivors had higher SP-A (from days 1 to 10 of symptoms onset) and lower CC16 (from days 11 to 20 of symptoms onset) levels vs survivors discharged from ICU. No significant differences in SP-D levels between the groups were found.Conclusion. According to the study results, the surfactant protein SP-A and Club cell protein CC16 are associated with increased COVID-19 mortality